microRNA介导TNF-α基因SNP在PCOS胰岛素抵抗中的机制

PCOS is the common reproductive endocrine metabolic disorder diseases. Local insulin resistance mediated by inflammatory factor in the ovary is the important pathological mechanism.However,the mechanism is not yet clear.Inflammatory factor and microRNA have complex regulation network. This project aims to study the role of tumor necrosis factor (TNF-α) gene SNP in PCOS-IR, and to make certain if microRNA mediates the TNF-α gene SNP function. In our previous studies, we confirmed that rs3179060C/T SNP sites of TNF-α was associated with PCOS.This study would systematically screen TNF-α gene SNP, and look for the SNP sites related with PCOS-IR;microRNA related with PCOS-IR would be analyzed using clinical biopsy by microRNA array;microRNA would be confirmed and then 1 or 2 SNP of PCOS-IR mediated by microRNA would be chosen.The biological functions on expression regulation would be further explored and the protein expression of IRS/PI3K/AKT/NFκB in the insulin signal pathway which is closely correlated insulin resistance,so as to provide new insights for the treatment and prevention of PCOS-IR.

多囊卵巢综合征(PCOS)是常见的生殖内分泌代谢紊乱疾病，由炎症因子引起的胰岛素抵抗(IR)是其重要病理环节。炎症因子与microRNA存在复杂的网络调控。本项目旨在研究肿瘤坏死因子α(TNF-α)基因SNP在PCOS-IR中的作用，明晰microRNA是否介导TNF-α基因SNP在胰岛素抵抗中起作用。我们前期研究证实TNF-αrs3179060C/T SNP位点与PCOS相关。本课题拟系统筛查TNF-α基因SNP多态位点，寻找与PCOS-IR相关SNP位点；以临床标本采用microRNA array、Northern blot等方法筛选并验证与PCOS-IR相关的microRNA,在此基础上筛选出1-2个受microRNA调控介导PCOS-IR的SNP并深入研究其对胰岛素信号通路IRS/PI3K/AKT/NFκB表达调控和生物学功能影响，为PCOS-IR防治提供新思路。

本项目研究了肿瘤坏死因子TNF-α基因rs3179060C/A，rs1799724C/T，rs361525G/A多态，雄激素受体AR基因CAG多态与PCOS的相关性，同时研究了不同基因型患者颗粒细胞TNF-α基因mRNA的表达与血清TNF-α浓度，颗粒细胞中的蛋白IRS1、NFκB和IκB表达研究。初步结果显示TNF-α基因rs3179060C/A多态与PCOS有一定的相关性，且具有A等位基因的患者血清的TNF-α的浓度明显升高，而rs1799724C/T，rs361525G/A多态与CAG多态未显示有相关性的趋势，但根据所含CAG的重复数,将其分为短组CAG(n≤22)与长组CAG(n>22)，PCOS患者组中短租率明显高于对照组。颗粒细胞中的TNF-α基因mRNA水平明显高于对照组。PCOS颗粒细胞中的IRS1、NFκB表达显著高于对照组，而IκB蛋白表达量显著低于对照组。这些结果提示TNF-α基因rs3179060C/A可能在炎症因子诱导的PCOS胰岛素抵抗中发挥了重要作用。